Functional genomic and molecular analyses of transcripts related to glycerol synthesis and trafficking in rainbow smelt (Osmerus mordax) using cold temperature-induced models of glycerol production

The rainbow smelt (Osmerus mordax) is an anadromous teleost that produces type II antifreeze protein (AFP) and accumulates modest urea and high glycerol levels in plasma and tissues as adaptive cryoprotectant mechanisms in sub-zero temperatures. It is known that glyceroneogenesis occurs in liver via a branch in glycolysis and gluconeogenesis and is activated by low temperature; however, the precise mechanisms of glycerol synthesis and trafficking in smelt remain to be elucidated. The objective of this thesis was to provide further insight using functional genomic techniques [e.g. suppression subtractive hybridization (SSH) cDNA library construction, microarray analyses] and molecular analyses [e.g. cloning, quantitative reverse transcription - polymerase chain reaction (QPCR)]. Novel molecular mechanisms related to glyceroneogenesis were deciphered by comparing the transcript expression profiles of glycerol (cold temperature) and non-glycerol (warm temperature) accumulating hepatocytes (Chapter 2) and livers from intact smelt (Chapter 3). Briefly, glycerol synthesis can be initiated from both amino acids and carbohydrate; however carbohydrate appears to be the preferred source when it is readily available. In glycerol accumulating hepatocytes, levels of the hepatic glucose transporter (GLUT2) plummeted and transcript levels of a suite of genes (PEPCK, MDH2, AAT2, GDH and AQP9) associated with the mobilization of amino acids to fuel glycerol synthesis were all transiently higher. In contrast, in glycerol accumulating livers from intact smelt, glycerol synthesis was primarily fuelled by glycogen degradation with higher PGM and PFK (glycolysis) transcript levels. Whether initiated from amino acids or carbohydrate, there were common metabolic underpinnings. Increased PDK2 (an inhibitor of PDH) transcript levels would direct pyruvate derived from amino acids and / or DHAP derived from G6P to glycerol as opposed to oxidation via the citric acid cycle. Robust LIPL (triglyceride catabolism) transcript levels would provide free fatty acids that could be oxidized to fuel ATP synthesis. Increased cGPDH (glyceroneogenesis) transcript levels were not required for increased glycerol production, suggesting that regulation is more likely by post-translational modification. Finally, levels of a transcript potentially encoding glycerol-3-phosphatase, an enzyme not yet characterized in any vertebrate species, were transiently higher. These comparisons also led to the novel discoveries that increased G6Pase (glucose synthesis) and increased GS (glutamine synthesis) transcript levels were part of the low temperature response in smelt. Glucose may provide increased colligative protection against freezing; whereas glutamine could serve to store nitrogen released from amino acid catabolism in a non-toxic form and / or be used to synthesize urea via purine synthesis-uricolysis. Novel key aspects of cryoprotectant osmolyte (glycerol and urea) trafficking were elucidated by cloning and characterizing three aquaglyceroporin (GLP)-encoding genes from smelt at the gene and cDNA levels in Chapter 4. GLPs are integral membrane proteins that facilitate passive movement of water, glycerol and urea across cellular membranes. The highlight was the discovery that AQP10ba transcript levels always increase in posterior kidney only at low temperature. This AQP10b gene paralogue may have evolved to aid in the reabsorption of urea from the proximal tubule. This research has contributed significantly to a general understanding of the cold adaptation response in smelt, and more specifically to the development of a working scenario for the mechanisms involved in glycerol synthesis and trafficking in this species.

Biological factors affecting sea survival of Atlantic salmon Salmo salar L., in Western Arm Brook and an analysis of selection by the commercial fishery of nearby St. Barbe Bay, Newfoundland

The problems faced by scientists in charge of managing Atlantic salmon (Salmo salar) stocks are : i) how to maintain spawning runs consisting of repeat spawners and large multi-sea-winter (MSW) adults in the face of selective homewater and distant commercial fisheries and , ii) how to more accurately predict returns of adults. Using data from scales collected from maiden Atlantic salmon grilse from two locations on the Northern Peninsula of Newfoundland, St. Barbe Bay and Western Arm Brook, their length at smolting was back calculated. These data were then used to examine whether the St. Barbe commercial fishery is selective for salmon of particular smolt age and/or size. Analysis indicated that come commercial fishery selected larger, but not necessarily older adults that those escaping to Western Arm Brook over the period of this study, 1978-1987. It was determined that less than average size smolts survived better than above average size smolts. Slection for repeat spawners, large MSW salmon, and larger grilse has meant reductions in the proportions of these adults in the spawning runs on Western Arm Brook. This may impact the Western Arm Brook salmon stock by increasing the population instability. Sea survival was significantly correlated with selection by the commercial fishery. Characteristics of adults in Western Arm Brook during the period of study (1978-1987) did not help in explaining yearly variation in sea survival. The characteristics of smolts, however, when subjected to multiple regression analysis explained 57.2 percent of the yearly variation in sea survival.

Glacial cold-water coral: ages, isotope concentrations and ratios

A set of 40 Uranium-series datings obtained on the reef-forming scleractinian cold-water corals Lophelia pertusa and Madrepora oculata revealed that during the past 400 kyr their occurrence in the Gulf of Cádiz (GoC) was almost exclusively restricted to glacial periods. This result strengthens the outcomes of former studies that coral growth in the temperate NE Atlantic encompassing the French, Iberian and Moroccan margins dominated during glacial periods, whereas in the higher latitudes (Irish and Norwegian margins) extended coral growth prevailed during interglacial periods. Thus it appears that the biogeographical limits for sustained cold-water coral growth along the NE Atlantic margin are strongly related to climate change. By focussing on the last glacial-interglacial cycle, this study shows that palaeo-productivity was increased during the last glacial. This was likely driven by the fertilisation effect of an increased input of aeolian dust and locally intensified upwelling. After the Younger Dryas cold event, the input of aeolian dust and productivity significantly decreased concurrent with an increase in water temperatures in the GoC. This primarily resulted in reduced food availability and caused a widespread demise of the formerly thriving coral ecosystems. Moreover, these climate induced changes most likely caused a latitudinal shift of areas withoptimum coral growth conditions towards the northern NE Atlantic where more suitable environmental conditions established with the onset of the Holocene.

(Table 2) Radiocarbon dating on cold-water corals collected in the Mediterranean Sea

This study presents newly obtained coral ages of the cold-water corals Lophelia pertusa and Madrepora oculata collected in the Alboran Sea and the Strait of Sicily (Urania Bank). These data were combined with all available Mediterranean Lophelia and Madrepora ages compiled from literature to conduct a basin-wide assessment of the spatial and temporal occurrence of these prominent framework-forming scleractinian species in the Mediterranean realm and to unravel the palaeo-environmental conditions that controlled their proliferation or decline. For the first time special focus was placed on a closer examination of potential differences occurring between the eastern and western Mediterranean sub-basins. Our results clearly demonstrate that cold-water corals occurred sparsely in the entire Mediterranean during the last glacial before becoming abundant during the Bølling-Allerød warm interval, pointing to a basin-wide, almost concurrent onset in (re-)colonisation after ~13.5 ka. This time coincides with a peak in meltwater discharge originating from the northern Mediterranean borderlands which caused a major reorganisation of the Mediterranean thermohaline circulation. During the Younger Dryas and Holocene, some striking differences in coral proliferation were identified between the sub-basins such as periods of highly prolific coral growth in the eastern Mediterranean Sea during the Younger Dryas and in the western basin during the Early Holocene, whereas a temporary pronounced coral decline during the Younger Dryas was exclusively affecting coral sites in the Alboran Sea. Comparison with environmental and oceanographic data revealed that the proliferation of the Mediterranean corals is linked with enhanced productivity conditions. Moreover, corals thrived in intermediate depths and showed a close relationship with intermediate water mass circulation in the Mediterranean sub-basins. For instance, reduced Levantine Intermediate Water formation hampered coral growth in the eastern Mediterranean Sea during sapropel S1 event as reduced Winter Intermediate Water formation did in the westernmost part of the Mediterranean (Alboran Sea) during the Mid-Holocene. Overall, this study clearly demonstrates the importance to consider region-specific environmental changes as well as species-specific environmental preferences in interpreting coral chronologies. Moreover, it highlights that the occurrence or decline of cold-water corals is not controlled by one key parameter but rather by a complex interplay of various environmental variables.

Regulatory factors controlling muscle mass : competition between innate immune function and anabolic signals in regulation of atrogin-1 in Atlantic salmon

The research was supported by an industrial PhD studentship between University of Aberdeen and by BioMar Ltd., for Z. Heidari.

Regulatory factors controlling muscle mass : competition between innate immune function and anabolic signals in regulation of atrogin-1 in Atlantic salmon

The research was supported by an industrial PhD studentship between University of Aberdeen and by BioMar Ltd., for Z. Heidari.

The trophic structure of Spongosorites coralliophaga-coral rubble communities at two northeast Atlantic cold water coral reefs

Funding for the JC073 cruise was provided by the Natural Environment Research Council (NERC) UK Ocean Acidification (UKOA) research programme’s Benthic Consortium project (NE/H017305/1 to J Murray Roberts). Funding for analytical costs and field work was provided by the Marine Alliance for Science and Technology Scotland (MASTS) (Biodiversity Grant to Ursula FM Witte, 140 SF10003-10). Georgios Kazanidis was funded by a MASTS PhD scholarship.

Constraints on community participation in salmon fisheries management in Northwest Russia

Acknowledgements Many parties contributed to making this paper a reality. This research was supported by the European Social and Research Council, grant ESRC ES/K006428/1. The author is particularly grateful to the grant’s holder, Professor David Anderson from the Department of Anthropology, University of Aberdeen, for his various support throughout this research. The Barents Center of the Humanities at Kola Science Center of the Russian Academy of Sciences in Apatity provided important institutional support. Officials from several fisheries management institutions of Arkhangelsk oblast, including Shiriaev Igor Alekseevich from Dvinsko-Pechorskoe Territorial Management Board, Skovorod’ko Artem Aleksandrovich from the Northern Basin Directorate of Fisheries and Water Biological Resources Conservation (Sevrybvod) and Korotenkov Aleksei Anatol’evich from the Fishing Industry Agency of Arkhangelsk oblast were very supportive and shared their knowledge wherever possible. Scholars Studenov Igor Ivanovich and Stasenkov Vladimir Aleksandrovich at Northern branch of the Knipovich Polar Research Institute of Marine Fisheries and Oceanography (SevPINRO) in Arkhangelsk provided their invaluable expertise on marine fisheries. Chairmen of several fishing collective farms – Tuchin Sergei Viktorovich, Samoilov Sergei Nikolaevich and Seliverstova Marina Nikolaevna – offered a great administrative support. Local residents of several villages in Mezen region were extremely generous and hospitable, providing places to stay, warm clothes, food, endless cups of tea, and most valuably, sparing their time. Finally, Natalie Wahnsiedler was a regular companion during fieldwork and a great source of inspiration for this research.

NFAT5 genes are part of the osmotic regulatory system in Atlantic salmon (Salmo salar)

Acknowledgements This study was supported by a grant from the Biotechnology and Biological Sciences Research Council (BBSRC, BB/H008063/1), UK to DGH and SAM. Funding also came from Research Council Norway for project number 241016 for DGH and EJ. This work was carried out as part of a PhD thesis funded by the Marine Alliance of Science and Technology Scotland (MASTS).

Constraints on community participation in salmon fisheries management in Northwest Russia

Acknowledgements Many parties contributed to making this paper a reality. This research was supported by the European Social and Research Council, grant ESRC ES/K006428/1. The author is particularly grateful to the grant’s holder, Professor David Anderson from the Department of Anthropology, University of Aberdeen, for his various support throughout this research. The Barents Center of the Humanities at Kola Science Center of the Russian Academy of Sciences in Apatity provided important institutional support. Officials from several fisheries management institutions of Arkhangelsk oblast, including Shiriaev Igor Alekseevich from Dvinsko-Pechorskoe Territorial Management Board, Skovorod’ko Artem Aleksandrovich from the Northern Basin Directorate of Fisheries and Water Biological Resources Conservation (Sevrybvod) and Korotenkov Aleksei Anatol’evich from the Fishing Industry Agency of Arkhangelsk oblast were very supportive and shared their knowledge wherever possible. Scholars Studenov Igor Ivanovich and Stasenkov Vladimir Aleksandrovich at Northern branch of the Knipovich Polar Research Institute of Marine Fisheries and Oceanography (SevPINRO) in Arkhangelsk provided their invaluable expertise on marine fisheries. Chairmen of several fishing collective farms – Tuchin Sergei Viktorovich, Samoilov Sergei Nikolaevich and Seliverstova Marina Nikolaevna – offered a great administrative support. Local residents of several villages in Mezen region were extremely generous and hospitable, providing places to stay, warm clothes, food, endless cups of tea, and most valuably, sparing their time. Finally, Natalie Wahnsiedler was a regular companion during fieldwork and a great source of inspiration for this research.

Regulatory factors controlling muscle mass : competition between innate immune function and anabolic signals in regulation of atrogin-1 in Atlantic salmon

The research was supported by an industrial PhD studentship between University of Aberdeen and by BioMar Ltd., for Z. Heidari.